Analysis of Cleavage Products of DNA Repair Enzymes and Other Nucleases. Characterization of an Apurinic/apyrimidinic Specific Endonuclease from Mouse Plasmacytoma Cells

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1988 Jul 25

PMID 2457203

Citations 2

Authors

B I Haukanes

D E Helland

K KLEPPE

Affiliations

Soon will be listed here.

Abstract

We have developed a strategy by which the nature of phosphodiester bond breaks produced by various DNA-repair endonucleases and also other nucleases, can be characterized. A purified apurinic/apyrimidinic (AP) specific endonuclease from a permanently established mouse plasmacytoma cell-line (MPC-11) has been examined with respect to the exact incision site generated at the baseless site. By the aid of enzymatic treatment with calf intestinal phosphatase, the 3'-phosphatase activity of T4-polynucleotide kinase, chemical modification with piperidine in addition to the Maxam-Gilbert sequencing procedure, followed by separation on a DNA-sequencing gel, the nature of the cleaved phosphodiester bond, both 3' and 5' to the cleavage site, has been established. The AP-specific endonuclease investigated was classified as a class II AP-endonuclease according to the four possible classes of AP-endonuclease with respect to the termini produced. By use of this technique each single damaged and cleaved site can be investigated separately.

Citing Articles

Mechanism of incision by an apurinic/apyrimidinic endonuclease present in human placenta.

Haukanes B, Wittwer C, Helland D Nucleic Acids Res. 1989; 17(14):5529-35.

PMID: 2474801 PMC: 318176. DOI: 10.1093/nar/17.14.5529.

Action of a mammalian AP-endonuclease on DNAs of defined sequences.

Haukanes B, Helland D, KLEPPE K Nucleic Acids Res. 1989; 17(4):1493-509.

PMID: 2466239 PMC: 331817. DOI: 10.1093/nar/17.4.1493.

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